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The changes of astaxanthin content and chemical characteristics of tiger

prawn (Penaeus monodon) due to processing: boiling, smoking and
frying
To cite this article: F Swastawati 2018 IOP Conf. Ser.: Earth Environ. Sci. 139 012050

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

The changes of astaxanthin content and chemical

characteristics of tiger prawn (Penaeus monodon) due to
processing: boiling, smoking and frying

F Swastawati
Fisheries Technology Study Program, Department of Fisheries, Faculty of Fisheries
and Marine Sciences, Diponegoro University, Jl. Prof. Soedarto, S.H., Tembalang,
Semarang, Central Java, Indonesia
*Email: fronthea_thp@yahoo.co.id

Abstract. Food processing using high temperatures can cause changes in pigment color and
chemical characteristics in food stuffs, including prawn. The aim of this research was to
evaluate the changes in pigment and chemical characteristics of tiger prawn caused by boiling,
smoking and frying. Ten kg of tiger prawn was boiled, smoked and fried at the temperature of
± 100 °C for ± 10 min. The results showed that boiling, smoking and frying gave a significant
effect (P < 0.05) on the astaxanthin pigment, pH, moisture, protein, salt content, Aw and color.
The content of astaxanthin pigments in fresh prawn, boiled prawn, smoked prawn and fried
prawn was: 132.79 ± 1.5 μg·g-1; 82.89 ± 0.92 μg·g-1; 78.28 ± 0.1 μg·g-1 and 91.35 ± 2.59 μg·g-
1
, respectively. The value of °Hue on fresh prawn, boiled prawn, smoked prawn and fried
prawn was: 87.85º; 52.5º; 55.94º and 53.98º. The tiger prawn processed by the smoking
method has preferable by panelist rather than processed by boiling and frying.

1. Introduction
Tiger prawn is a popular seafood in many countries and it becomes a favorite seafood choice by
consumers because of its specific taste and aroma. Although it has a tangy smell in its raw condition,
after processing, distinct flavor and specific delicious taste will arise. Based on data from the Ministry
of Marine Affairs and Fisheries in 2013, Indonesia exported shrimp to various destination countries
reaching a total volume of 162,000 tons. Production of shrimp aquaculture during the last five years
(2010–2014) has a tendency to continue rising. The average increase of shrimp production during the
last five years is 13.83 % per year. In 2012 shrimp production of 415,703 tons was reached while in
2013 amounted to 645,955 tons [1].
Food processing by heating which is also known as the cooking process is the process of heating
food at a temperature of 100 °C or more. The main purpose is to get a better taste, better aroma and
softer texture; moreover, it also kills microbes and deactivates all enzymes. Cooking can be done by
boiling, smoking and frying. The use of heat in the cooking process is very influential on the
nutritional value of the foods including boiling, smoking and frying. The goals of the salt-boiled
process for shrimp are to reduce a load of microorganisms to an acceptable level and to improve the
flavor of shrimp. A suitable boiling process is important to get a suitable condition to minimize loss of
muscle protein and maintain the red color. Smoking by using liquid smoke can produce smoked fish
that is higher in quality when compared with the traditional smoking process. Smoking by using liquid
smoke may affect the color and sensory quality of smoked fish products [2]. Frying is a fast and
simple culinary technique employed worldwide that consists of cooking food above the boiling point

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

of water, by means of partial (shallow-frying) or total (deep-frying) immersion in liquid edible oils or
fats at frying temperature. Although fried foods are highly appreciated for their sensory characteristics,
they must also satisfy health-related consumer concerns.
High-temperature processing can also affect the change of pigment and chemical characteristics in
foodstuffs, including prawn. Prawn color turns red because it contains astaxanthin, a pigment that
gives the characteristic of orange to red. Changes in pigment can also indicate that there has been a
change in chemical characteristics of prawn. The purpose of this research was to evaluate the effect of
processing treatments on the changes of astaxanthin content and chemical characteristics in tiger
prawn.

2. Materials and Methods

2.1. Materials
The material used in this study was ± 10 kg of tiger prawn with a size of ± 20 cm in length and an
average weight of ± 150 g. Cooking oil, the food grade liquid smoke made of coconut shell, boiling
water and salt as a flavor enhancer.

2.2. Methods
This research was conducted experimentally using completely randomized design with duplication.
After washed thoroughly, the prawns were dipped into 5 % liquid smoke and 5 % of brine solution.
Then they were processed by boiling (at ± 100 °C), smoking (at ± 100 °C) and frying (at ± 100 °C) for
± 10 min [3].

2.3. Measurement of astaxanthin content

Astaxanthin content was measured following Becerra et al. [4], a total of 10 g samples was extracted
with 40 mL 0.05 % BHT solution (in acetone) with a digital homogenizer for 2 min. The samples were
cooled during the homogenization process to prevent the sample from heating up. After extraction, the
sample was centrifuged at 4,000 rpm, temperature of 40 ºC for 5 min. To separate the water-soluble
compound, the acetone extract from the sample was transferred to a 250 mL separating funnel with 40
mL n-hexane. Next, 100 mL of aquadest containing 0.5 % (w/v) of sodium chloride was added to the
mixture. After continuous shaking, phase separation was achieved and then the top layer was separated
and transferred into 50 mL measuring flask. The absorbance spectrum of a water-soluble compound
was measured at a wavelength of 472 nm using a spectrophotometer. The astaxanthin stock solution
(0.2 mg·mL-1) of 0.5; 1.0; 1.5; 2.0 and 3.0 mL were respectively diluted with 10 mL n-hexane. Then
the absorbance spectrum of each solution was recorded with a wavelength of 472 nm. The standard
curve was obtained by plotting the concentration on absorbance.

2.4. Measurement of pH
The sample was ground and weighed as much as 1 g in a cup glass. Then, 10 mL of aquadest was
added and stirred. Subsequently, pH level of the sample in the container was measured by using a
calibrated pH meter with a buffer solution of pH 4 and pH 7. The pH value was obtained based on the
reading at pH meter until the digital number indicated the constant number [5].

2.5. Measurement of moisture content

The determination of moisture content was based on the different weight of samples before and after
drying. The method used was a drying technique for approximately 24 h at 105 °C until the sample
reached a stable weight. Percentage of moisture contents (wet weight) was calculated by using the
following formula [5]:
B-C
Moisture Content (%)= x 100 %
B-A

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

Information:
A: empty vial weight (g)
B: cup weight + initial sample (g)
C: cup weight + dry sample (g)

2.6. Measurement of protein content

Determination of protein content was done based on AOAC [5]. Basically, it could be divided into
three stages namely the process of destruction, distillation and titration. In the process of destruction
the sample was heated with concentrated H2SO4 thus it broke down into their respective elements. The
destruction process was completed when the solution became clear. The distillation stage resulted in
the broken down of ammonium sulfate into ammonia by adding alkalis NaOH and heated. The formed
ammonia which was accommodated in concentrated H 3BO3 added with BCG and methyl red indicator.
The amount of H3BO3 reacted with ammonia could be known by its reaction with 0.02 M HCl. The
end of the titration was marked by the change of solution color from green to pink. Blank treatment
was performed to determine the nitrogen from the reagents being used.

2.7. Measurement of Aw
The instrument was calibrated by inserting the BaCl2 2H2O liquid and closed for 3 min to reach the
reading scale of 0.9. Then, samples were inserted into Aw meter for 3 min and Aw scale was recorded.
If the temperature scale reached above 20 °C, the scale reading was added as much as the excess
temperature multiplied by the correction factor of 0.002 °C, as well as the temperature below 20 °C
[6].

3. Results and Discussion

3.1. Astaxanthin content

Table 1. Astaxanthin content tiger prawn before and after the heating
process.
Treatment Astaxanthin (μg·g-1)
Fresh 132.79 ± 1.5a
Boiled 82.89 ± 0.92b
Smoked 78.28 ± 0.1c
Fried 91.35 ± 2.59d
Note: Value from average of duplicate ± standard deviation.
*Value with different lower case notation shows a significant difference (p < 0.05).

Table 1 shows that the content of astaxanthin in tiger prawn decreased after heat processing. The
heating processes led to the reduction of astaxanthin levels as the proteins protecting astaxanthin had
been denatured by heat, resulting in the decomposition of astaxanthin. In boiling treatment, this
reduction could be related to denaturation and partial solubilization of the carotene protein complex. In
this regard, a significant decrease in proteins content (myofibrillar, sarcoplasmic and stroma) in boiled
shrimp, suggesting that part of these proteins could have been dissolved in the salt solution [7]. The
reduction of astaxanthin due to heat is also reported in drying treatment. Astaxanthin content in
cooked shrimp decreased by 75 % after four days of direct sun drying, reaching the concentration of
25.0 ± 2.7 µg·g-1 [4]. The authors noted that the decline in astaxanthin content in shrimp during sun
drying mainly affects the color of the product.

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

3.2. Moisture content

Table 2. Moisture content, protein content and water activity of tiger prawn
before and after the heating process.
Treatment Moisture content (%) Protein content (%) Aw
Fresh 88.72 ± 0.54a 27.6 ± 1.5a 0.93 ± 0.0007a
Boiled 72.7 ± 0.32b 14.17 ± 1.4b 0.83 ± 0.0007b
c c
Smoked 59.84 ± 0.59 22.57 ± 0.23 0.72 ± 0.0014c
d d
Fried 57.6 ± 1.17 14.3 ± 0.68 0.62 ± 0.002d
Note: Value from average of duplicate ± standard deviation.
*Value with different lower case notation shows a significant difference (p < 0.05).

The lowest moisture content was found in fried prawn at 57.6 ± 1.17 % with a reduction of 35.076 %,
followed by the smoked prawns with a reduction of 32.55 %, and boiled prawn with a reduction of
18.05 %. The reduction in water content as a consequence of processing related to the temperature
being used, the higher the temperature the higher the water content decreases. It happened because of
the process of steaming and boiling used temperature of 90–100 °C, while the frying process used a
temperature of 200–205 °C. As a comparison study by previous researchers [8], showed that the
largest reduction in water content occurred in soybean cake (20.13 %) followed by chicken (16.74 %),
mackerel (16.05 %) and the smallest was in tofu (7.66 %).
Moisture content is related to the presence of microorganisms that grow in the body of prawn since
water is a medium that supports the growth of microorganisms. The lower the water content, the lower
microorganisms decompose. Water is the largest content in fish [9]. Water is a means of
microorganisms to thrive. The moisture content reduction in fish can extend the shelf life of the
products. Salting process is capable oflosing water in fish because salt has a higher concentration than
water, so the water will come out of the tissue or called plasmolysis. The decrease of the moisture
content was the most prominent change in fish fillet after drying. Protein, fat and ash content increased
significantly in the processed sample [10].

3.3. Protein content

Protein content in prawn decreased after treated with boiling, smoking or frying process. Protein
content in both boiled and fried prawn had almost the same value of 14.17 ± 1.4 % and 14.3 ± 0.68 %,
respectively, with a decrease of 48.19 %. The lowest reduction in protein content occurred in smoked
prawn (18.22 %) with a protein content of 22.57 ± 0.23 %. The reduction of protein content was
directly proportional to the used processing temperature, meanwhile, in the process of frying and
boiling the used temperature was more than 100 °C hence the protein was damaged by heat. Other
researchers reported that the reduction of protein content in boiled bloated fish was 3.12 % whereas
fried bloated fish was 5.24 % [8]. During cooking, heat induces protein denaturation and aggregation,
causing actin, myosin and collagen shrinkage, which leads to water expulsion from muscle cells and
the consequent water loss. During boiling, protein content also decreased. Water and protein losses are
called cooking loss [7].
Processing with high temperatures can lead to proteins damage because the protein bonds have
undergone changes and breaks. It also deals with the solubility of proteins, water-soluble proteins and
faster salts lost due to heat. Protein solubility decreased with increasing temperature, it indicated
changes in protein solubility [3]. Denaturation of proteins related to protein solubility, as a result of the
heat, the secondary or tertiary structures of the protein was damaged, hence it transformed into a
primary structure. Salt-soluble and water-soluble proteins were more easily lost in the primary
structure rather than those in secondary or tertiary structures. Boiling, smoking and frying maintained
and concentrated the crude protein content. After storage, there was a reduction in crude protein
content of the fried fish rather [11].

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

3.4. Water activity

Aw of fresh prawn was 0.93. The lowest Aw occurred in fried prawn with a value of 0.62.While the Aw
of smoked and boiled prawns were 0.72 and 0.83, respectively. Processing with high temperatures
could decrease the value of Aw. It was in line with the decrease in water content. The relationship of
water content with water activity (Aw) was indicated by the tendency that the higher the water content
the higher the Aw value would be and vice versa [12].

Table 3. The value of pH and salt content of tiger prawn before and after
the heating process.
Treatment pH Salt content (%)
Fresh 7.6 ± 0.14a 0.78 ± 0.04a
b
Boiled 7.55 ± 0.07 1.4 ± 0.16b
c
Smoked 6.45 ± 0.07 4.2 ± 0.12c
d
Fried 7.3 ± 0.14 3.4 ± 0.07d
Note: Value from average of duplicate ± standard deviation.
*Value with different lower case notation shows a significant difference (p < 0.05).

3.5. pH value
The pH value of all samples decreased from the initial pH of fresh prawn with a value of 7.6. The
value of pH on boiled prawn and fried prawn were 7.55 and 7.3, respectively. The lowest pH value
occurred in smoked prawns with a value of 6.45. The pH value in smoked prawn was acidic, whereas
the pH value of other treatments indicated that the pH was neutral. The low pH of the smoked prawns
occurred as the effect of the addition of liquid smoke prior to processing. This acidic pH trend was
caused by the absorption of the liquid organic liquids into the sample. The longer the immersion of
liquid smoke, the more organic acids such as acetic acid, formic acid would enter into the fish meat
during smoking process and resulted in a reduction of pH value. The smoke attached to the fish meat
caused the fish meat to be more acidic therefore the pH value tends to be lower [13].

3.6. Salt content

Samples were dipped into 5 % of brine before heat processing. Salt content in all samples decreased
significantly after processing. The decrease in salt content was caused by salt dissolution into
processing media such as oil and water. The highest salt content was found in smoked prawns with a
value of 3.4 %, while the lowest was found in boiled prawn with a value of 1.4 %. The salt content of
smoked catfish ranges from 2.62 to 5.83 % [3]. In this study, cooking loss observed was 32 %, which
is higher than losses reported by previous researchers [7] for boiled shrimp in a salt solution (4 %,
w/v) for 7 min. The greater cooking loss we found can be attributed to the longer boiling time (15 min
vs. 7 min). On the other hand, the NaCl content in shrimp showed a significant increase after boiling
in brine.

3.7. Colour analysis

Processing affected the color change in prawn. The value of °HUE of boiled prawn reduced 35.35
°HUE into the yellow-redcolor category, the smoked prawn reduced 31.91 °HUE and fried prawn
reduced 33.87 °HUE into the red color category (table 4). Changes in astaxanthin are marked in red
color on prawn. Colour changes from reddish pink to orange in prawn are formed by the release of
astaxanthin from protein-bound due to proteolysis [14].

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

Table 4. Colour analysis of tiger prawn before and after the heating process.
Colour
°
Treatment L +a* +b* HUE
(Lightness) (Redness) (Yellowness)
Fresh 35.64 ± 0.7a 0.2 ± 0.01a 5.32 ± 0.66a 87.85d
c c c
Boiled 46.57 ± 0.49 14.77 ± 0.68 19.23 ± 0.71 52.5b
e e e
Smoked 48.24 ± 0.22 15.63 ± 0.44 23.12 ± 0.08 55.94a
b a b
Fried 41.32 ± 0.33 12.25 ± 0.45 16.85 ± 1.75 53.98f
Note: Value from average of duplicate ± standard deviation.
*Value with different lower case notation shows a significant difference (p < 0.05).

3.8. Sensory value of fresh prawn

The results of thesensory value of fresh prawn showthat appearance, odor, and texture covered average
values of 8.73 ± 0.45; 8 ± 0.51 and 8.8 ± 0.41, respectively. Fresh prawn had a specific prawn’s
texture of elastic, compact and solid. Based on these specifications, the prawns used in this research
were still in the fresh category. The sensory of fresh prawn fulfill requirements of sensory of fresh fish
based on Indonesian National Standard for fresh fish which minimal sensory value was seven.

3.9. The prawn preferences test

Table 5. Preferences test of tiger prawn after heat processing treatment.

Hedonic parameter
Treatment
Appearance Flavour Odor Texture
Boiled prawn 6.5 ± 0.51a 6.6 ± 0.56a 7.2 ± 0.48a 6.23 ± 0.5a
Smoked prawn 8.57 ± 0.5c 8.1 ± 0.31b 8 ± 0.49b 8.57 ± 0.5c
b b a
Fried prawn 7.57 ± 0.5 8.1 ± 0.42 7.3 ± 0.48 7.87 ± 0.57b
Note: Value from anaverage of 10 panelists ± standard deviation.
*Value with different lowercase notation shows a significant difference (p < 0.05).

3.9.1. The appearance. The panelists gave the highest score for the appearance of the smoked prawn.
The results of the appearance were as follows: surface quality prawn was bright, shiny, invisible dirt in
the form of dried blood and the specific color of prawn. Chemical reactions among substances
contained in the smoke could occur in smoked fishery products. The chemical reaction between
phenol and oxygen could change the color of the smoke product to be a brownish yellow. Phenol, the
major carbonyl component in smoke reacting with proteins and fats in fish, has an important role in
affecting the appearance of smoked fish [9].

3.9.2. Flavors. The panelist's preference score in the flavor of both smoked and fried prawn was the
highest with a similar average value of 8.1. The taste specifications on fried prawn were savory on the
contrary the smoked prawn had a distinctive taste of smoke products. In addition, boiled prawn had a
slightly bland taste when compared with the other two processing treatments. The acceptance of
panelists on taste parameters was based on not only the maturity level but also taste of “umami” in the
processed prawn. Processing techniques with heating were able to produce products that have
extraordinary taste compared with other techniques [15].

3.9.3. Odor. The highest panelists preference score of the odor was in the smoked prawn based on the
specification that they had the distinctive aroma of smoke. It was the effect of the increase of phenol
levels due to heating. The increase of phenol level occurred because of temperature and heating time.
The higher the temperature and curing time resulted to the higher the phenol production [9]. In fact,
the aroma of boiled prawn and fried prawn was still included in a favorite category with a value of 7.2

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The 2nd International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 139 (2018)
1234567890 ‘’“” 012050 doi:10.1088/1755-1315/139/1/012050

± 0.48 and 7.3 ± 0.48. The aroma of a food that undergone high-temperature processing was caused by
the volatile compounds. The aroma produced in food after processing affected by the compound in
food and the way of cooking [15].

3.9.4. Texture. The highest score for texture preference was in the smoked prawns with a value of
8.57 ± 0.5. The texture of smoked prawn was solid, compact and not fragile. The addition of liquid
smoke might affect the texture of prawn result from the reaction of liquid smoke components such as
phenol. The main carbonyl component in the smoke played an important role. The smell, taste and
texture of smoked fish were the results of the reaction of carbonyl groups in the smoke reacting with
proteins and fats in fish. Smoke plays an important role in the formation of color, texture and taste [9].

4. Conclusion
The different prawn processing by boiling, smoking or frying affected pigment concentration changes
and chemical characteristics in tiger prawn. Further research on the influence of various types of oil in
frying, various types of liquid smoke and different boiling temperature on the changes in pigment
concentration and chemical characteristics of prawn needs to be conducted in the future.

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